Fig. 1: Synthetic lethality screen using engineered ovarian cancer cells.
From: Synergistic targeting of cancer cells through simultaneous inhibition of key metabolic enzymes
A Upper: iFTSECs were transduced with a lentivirus encoding KRASG12V and MYC oncogenes separated by the porcine teschovirus-1 2 A (P2A) sequence to allow for polycistronic gene expression or the empty vector as a control. Lower: Three months after transduction and selection, proliferation was analyzed for 12 days. Shown are mean ± SD, normalized to t = 0 days, n = 3, multiple t test, FDR q values. B Left: KRASG12V/MYC cells and EV control cells were stained for Actin and Tubulin and examined for cytoskeletal architecture and cell area by immunofluorescence microscopy. Scale bars = 50 µm. Right: Cell-area quantification of 51 cells, violin plots are shown and statistics are presented as two-tailed, unpaired t-test. C 2 ×104 of the indicated cells were seeded in soft agar and stained with crystal violet after 9 days of growth. D Extracellular acidification rate (ECAR) of the indicated cells was analyzed by Seahorse metabolic flux analysis, maximal ECAR was determined in the presence of oligomycin (2 µM). Shown are mean ± SD, n = 4, two-way ANOVA with Šídák’s multiple comparisons test. E Sublethal concentrations of the indicated metabolic inhibitors (Supplementary Table 1) were added to KRASG12V/MYC cells in 96-well plates. After 3 days, cell densities were determined by staining with crystal violet and quantification in a plate reader. F The indicated combinations of Metformin, GNE and BMS were added to the cells. After 3 days, cell densities were quantified by crystal violet staining, data are shown as mean ± SD, n = 3, two-way ANOVA with Šídák’s multiple comparisons test. G KRASG12V/MYC and EV control cells were treated with GNE and/or BMS for 3 days followed by determination of cell densities by crystal violet staining. Shown are mean ± SD, n = 5, two-way ANOVA with Dunnett’s and Šídák’s multiple comparisons test.
